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Dovyalis Hebecarpa) Pulp ⇑ Vivian Caetano Bochi A, , Milene Teixeira Barcia A, Daniele Rodrigues A, Caroline Sefrin Speroni B, M

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Dovyalis Hebecarpa) Pulp ⇑ Vivian Caetano Bochi A, , Milene Teixeira Barcia A, Daniele Rodrigues A, Caroline Sefrin Speroni B, M View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Food Chemistry 164 (2014) 347–354 Contents lists available at ScienceDirect Food Chemistry journal homepage: www.elsevier.com/locate/foodchem Analytical Methods Polyphenol extraction optimisation from Ceylon gooseberry (Dovyalis hebecarpa) pulp ⇑ Vivian Caetano Bochi a, , Milene Teixeira Barcia a, Daniele Rodrigues a, Caroline Sefrin Speroni b, M. Monica Giusti c, Helena Teixeira Godoy a a Department of Food Science, Faculty of Food Engineering, Campinas State University (UNICAMP), P.O. Box 6121, 13083-862 Campinas, SP, Brazil b Integrated Center for Laboratory Analysis Development (NIDAL), Department of Food Technology and Science, Center of Rural Sciences, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil c The Ohio State University, Department of Food Science and Technology, 2015 Fyffe Road, Columbus, OH 43210, United States article info abstract Article history: Originally from Asia, Dovyalis hebecarpa is a dark purple/red exotic berry now also produced in Brazil. Received 27 October 2013 However, no reports were found in the literature about phenolic extraction or characterisation of this Received in revised form 8 May 2014 berry. In this study we evaluate the extraction optimisation of anthocyanins and total phenolics in Accepted 11 May 2014 D. hebecarpa berries aiming at the development of a simple and mild analytical technique. Multivariate Available online 16 May 2014 analysis was used to optimise the extraction variables (ethanol:water:acetone solvent proportions, times, and acid concentrations) at different levels. Acetone/water (20/80 v/v) gave the highest anthocyanin Keywords: extraction yield, but pure water and different proportions of acetone/water or acetone/ethanol/water D. hebecarpa (with >50% of water) were also effective. Neither acid concentration nor time had a significant effect Response surface methodology Simplex lattice design on extraction efficiency allowing to fix the recommended parameters at the lowest values tested Anthocyanin (0.35% formic acid v/v, and 17.6 min). Under optimised conditions, extraction efficiencies were increased Phenolic by 31.5% and 11% for anthocyanin and total phenolics, respectively as compared to traditional methods Extraction optimisation that use more solvent and time. Thus, the optimised methodology increased yields being less hazardous and time consuming than traditional methods. Finally, freeze-dried D. hebecarpa showed high content of target phytochemicals (319 mg/100 g and 1421 mg/100 g of total anthocyanin and total phenolic content, respectively). Ó 2014 Elsevier Ltd. All rights reserved. 1. Introduction the mouth due to a velvety texture and could be considered co- product (Morton & Dowling, 1987). Recently, it is being cultivated Ceylon gooseberry or Ketembilla (Dovyalis hebecarpa, Salicaceae as an exotic fruit in the southwest regions of Brazil. High produc- family), originally from Sri Lanka and introduced into USA (Florida) tion is obtained from March to May, but it is cultivated until in 1920, is a small spherical fruit (0.5–1 inch) characterised by a August with satisfactory harvesting yields. deep purple-red colour, sour juice, and small seeds enclosed in A hybrid of D. hebecarpa with Dovyalis abyssinica was reported the pulp. Fruits can be consumed as jams, juice, or fresh after skin as a source of vitamin C (120.3 mg/100 g of fresh fruit) and with removal. Despite an attractive appearance, skins are unpleasant in good physical quality for market, showing an average of 75% pulp (Cavalcante & Martins, 2005). Phenolic composition of this hybrid revealed higher contents of anthocyanins in fruit peels and carote- Abbreviations: SLD, simplex lattice design; RSM, response surface methodology; noids in the pulp (De Rosso & Mercadante, 2007). Nevertheless, no TMA, total monomeric anthocyanin; TPC, total phenolic content; GAE, gallic acid reports were found about extraction and phytochemicals content equivalent; CGE, cyanidin-3-glucoside equivalent. in D. hebecarpa species. ⇑ Corresponding author. Address: Santa Maria Federal University (Universidade Federal de Santa Maria, UFSM), Rural Science Center (Centro de Ciências Rurais, Phenolic compounds are secondary metabolites in fruits and CCR), Food Technology and Science Department (Departamento de Tecnologia e vegetables acting as a defense barrier against microorganism, Ciência de Alimento, DTCA), 42 Building, 97119-900 Santa Maria, RS, Brazil. Tel.: insects, and UV radiation, or as attractants to promote pollination +55 55 3213 4890; fax: +55 19 3521 2153. and seed dispersal. Some phytochemicals, as anthocyanins and E-mail addresses: [email protected] (V.C. Bochi), [email protected] phenolic acids, are very important for food acceptance acting as (M.M. Giusti), [email protected] (H.T. Godoy). http://dx.doi.org/10.1016/j.foodchem.2014.05.031 0308-8146/Ó 2014 Elsevier Ltd. All rights reserved. 348 V.C. Bochi et al. / Food Chemistry 164 (2014) 347–354 natural pigments or adding specific flavours, such as astringency Due to the increasing need for efficient, simple, and mild and acidity (Shahidi & Naczk, 2003; Lattanzio, Kroon, Quideau, & extraction procedures for anthocyanins and other phenolics to Treutter, 2009). allow its further evaluation and characterisation, this work was Potential health benefits of phenolic compounds in the human focused on optimising extraction methods for these types of diet have been extensively discussed in the literature and are still compounds from D. hebecarpa pulp. Within this study, multi- being investigated mainly linked to antioxidant activity and variate experimental designs were used to develop procedures degenerative disease prevention (Zafra-Stone et al., 2007). The bio- investigating extraction variables to improve knowledge in this logical activity of berry phytochemicals is believed to be a result of field. multiple mechanisms that initially were believed to be mainly linked to antioxidant effects. However, nowadays the questionable 2. Material and methods polyphenols bioavailability has given rise to the discussion of different mechanisms by which it could act in the onset and in the 2.1. Chemicals and equipment development of degenerative diseases. Some of them are their anti-inflammatory activity, their enzyme inhibition capacity, The phenol reagent Folin–Ciocalteu and gallic acid were or their potential modulation of the gut microbiota (Chiva- obtained from Sigma–Aldrich. A Büchi rotary evaporator and an Blanch & Visioli, 2012). UV-1600 spectrophotometer from Proanálise (São Paulo, Brazil) Anthocyanins are glycosylated forms of polyhydroxy and were used for concentration and quantification purposes. A food polymethoxy derivatives of 2-phenylbenzopyrylium, acylated or processor (Philips Mini Food Processor, model HR7625) Terroni not, which are found as red to purple natural pigments in fruit Freeze-dryer, model LS-3000E (São Carlos, Brazil) and a Qhimis and vegetables. Usually, extraction procedures are conducted using analytical grinder, model Q298A (São Paulo, Brazil) were used for acidified solvents due to the higher anthocyanin stability at low pH sample preparation. values (Giusti & Jing, 2008). However, degradation of native chemical structure due to acid hydrolysis or oxidation can occur if 2.2. Sample preparation the acid concentration is high or if the extraction time is long (Naczk & Shahidi, 2006; Revilla, Ryan, & Martín-Ortega, 1998). Ripened samples (12.5 °Brix) were obtained from Bragança Thus, extraction conditions should be studied to ensure high Paulista city (São Paulo, Brazil) in April of 2010 (see Supplementary extraction yields and minimal degradation in native chemical material for fruit images). After harvesting, fruits were manually structure of these pigments. Finally, results obtained under peeled while frozen (À20 °C) to minimise enzymatic degradation optimised extraction conditions will better represent the real com- and decrease juice loss. Frozen pulps were crushed using a food position of the vegetable matrix being used for characterisation processor and placed into trays to re-freeze. Materials were then studies. freeze-dried until the pressure was reduced to stable values lower Aiming to develop efficient and fast new extraction techniques, than 22lHg. Since particle size is extremely relevant to extraction sonication and subcritical fluids have been tested with promising effectiveness, freeze-dried samples were grinded until a fine and results (Adil, Cetin, Yener, & Bayındırlı, 2007). However, these flu- visually homogenous powder was obtained. ids increase the extraction costs and require equipment that is not typically present in most laboratory facilities. Moreover, in some cases, traditional methods still give better yields. Previous litera- 2.3. Initial extraction procedure ture data revealed that solvent extraction showed higher results A water/acetone (25:75 v/v) mixture, acidified with 2% of formic than an optimised subcritical fluid method (CO2 plus ethanol) in apple and peach pomace for total phenolics and antioxidant activ- acid was the initial solvent tested (Mertz, Cheynier, Günata, & Brat, ity (Adil et al., 2007). Nevertheless, the presence of even relatively 2007). Freeze-dried powder samples (0.5 g) were mixed with the safer solvents (such as alcohol) in the composition of natural colo- extraction solvent (15 ml) in a ratio of 1:30 w/v for 15 min, filtered rants may limit its application
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